Nucleus transfer in mammals: noninvasive approaches for the preparation of cytoplasts

Fulka, Josef; Loi, Pasqualino; Fulka, Helena; Ptak, Grazyna; Nagai, Taku

doi:10.1016/j.tibtech.2004.04.002

Cited by 42 publications

(19 citation statements)

References 33 publications

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“…The previous report has indicated that in the event of a functional failure of the spindle, the first polar body was not extruded (32). On the contrary, the very similar phenotype of oocytes of the present study, a completely aggregated one chromosome mass in the polar body with decreased MPF activity, has been reported after etoposide‐cycloheximide, topoisomerase II inhibitor‐protein synthesis inhibitor, respectively, a treatment for noninvasive enucleation of mouse MII oocytes for animal cloning (33, 34). Therefore, our observation of morphologically normal polar bodies after the loss of Obox4 and the accompanying abnormalities in spindle and chromosome organization with decreased level of MPF strongly suggests that Obox4 is critical in the regulation of MI‐MII transition and cytokinesis during meiosis.…”

Section: Discussionsupporting

confidence: 84%

Obox4 critically regulates cAMP‐dependent meiotic arrest and MI‐MII transition in oocytes

Lee¹,

Kim²,

Kim³

et al. 2010

FASEB j.

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Extra follicular oocytes spontaneously resume meiosis in vitro, but the intact germinal vesicle (GV) is retained if the oocytes are cultured in medium containing phosphodiesterase (PDE) inhibitors or cAMP analogues. On the basis of our finding that Obox4 is prominently expressed in oocytes, the present study was conducted to determine the functional role of the homeodomain-containing factor Obox4 during in vitro oocyte maturation. After microinjection of Obox4 dsRNA into the cytoplasm of GV oocytes cultured in M16 medium, oocytes were arrested at metaphase I (MI, 77.7%) and metaphase II (MII, 22.3%). Surprisingly, however, 89% of Obox4 RNAi-treated oocytes resumed meiosis and developed to MI and MII when cultured in medium containing 0.2 mM 3-isobutyl-1-methyl-xanthine (IBMX), in which untreated oocytes maintain intact GVs. Spindles were aberrant, and chromosomes were severely aggregated with decreased MPF and MAP kinase activities in arrested MI oocytes after exposure to Obox4 RNAi. Oocytes overexpressing Obox4 retained intact GVs when cultured in M16 medium. Taken together, for the first time to our knowledge, these findings indicate that Obox4 plays a key role in the cAMP-dependent signaling cascades that maintain GV arrest. Oocytes not expressing Obox4 failed to maintain intact GVs in IBMX-supplemented medium, while GVs remained intact when oocytes were kept in plain medium and overexpressing Obox4, suggesting that Obox4 plays a critical role in cAMP-dependent cascade for maintaining intact GVs.

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Section: Discussionsupporting

confidence: 84%

Obox4 critically regulates cAMP‐dependent meiotic arrest and MI‐MII transition in oocytes

Lee¹,

Kim²,

Kim³

et al. 2010

FASEB j.

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“…Several approaches have been developed in order to improve and facilitate oocyte enucleation (reviewed in Fulka et al, 2004;Li et al, 2004). Chemical enucleation using microtubule depolymerizing drugs is a very attractive procedure that could simplify conventional enucleation methods and that avoids the removal of the meiotic spindle.…”

Section: Introductionmentioning

confidence: 99%

Antimitotic Treatments for Chemically Assisted Oocyte Enucleation in Nuclear Transfer Procedures

Costa-Borges

Paramio

Calderón

et al. 2009

Cloning and Stem Cells

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Chemically assisted enucleation has been successfully applied to porcine and bovine oocytes to prepare recipient cytoplasts for nuclear transfer procedures. In this study, the antimitotic drugs demecolcine, nocodazole, and vinblastine were first assessed for their ability to induce the formation of cortical membrane protrusions in mouse, goat, and human oocytes. While only 2% of the treated human oocytes were able to form a protrusion, high rates of protrusion formation were obtained both in mouse (84%) and goat oocytes (92%), once the treatment was optimized for each species. None of the antimitotics applied was superior to the others in terms of protrusion formation, but mouse oocytes treated with vinblastine were unable to restore normal spindle morphology after drug removal and their in vitro development after parthenogenetic activation was severely compromised, rendering this antimitotic useless for chemically assisted enucleation approaches. Aspiration of the protrusions in mouse oocytes treated with demecolcine or nocodazole yielded 90% of successfully enucleated oocytes and allowed the extraction of a smaller amount of cytoplasm than with mechanical enucleation, but both enucleation methods resulted in the depletion of spindle-associated gamma-tubulin from the prepared cytoplasts. Treatment of mouse oocytes with demecolcine or nocodazole had no effect on their in vitro development after parthenogenetic activation, or on their ability to repolymerize a new spindle after the removal of the drug or the reconstruction of the treated cytoplasts with a somatic nucleus. Therefore, demecolcine- and nocodazole-assisted enucleation appears as an efficient alternative to mechanical enucleation, which can simplify nuclear transfer procedures.

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“…The pre-implanted embryos are then maintained in a sequential culture media to support development. Finally the developed embryo is transferred to a foster mother (Fulka, Loi, Fulka, Ptak, & Nagai, 2004). Various somatic cells, including mammary epithelial cells, cumulus cells, oviductal cells, leukocytes, hepatocytes, epithelial cells, neuronal cells, myocytes, lymphocytes and germ cells have been successfully used as donor cells for production of cloned animals (Brem & Kuhholzer, 2002; Hochedlinger & Jaenisch, 2002; Jaenisch, et al, 2004).…”

Section: Techniques To Generate Pluripotent Stem Cellsmentioning

confidence: 99%

Advances in Reprogramming Somatic Cells to Induced Pluripotent Stem Cells

Patel

Yang

2010

Stem Cell Rev and Rep

183

128

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Traditionally, nuclear reprogramming of cells has been performed by transferring somatic cell nuclei into oocytes, by combining somatic and pluripotent cells together through cell fusion and through genetic integration of factors through somatic cell chromatin. All of these techniques changes gene expression which further leads to a change in cell fate. Here we discuss recent advances in generating induced pluripotent stem cells, different reprogramming methods and clinical applications of iPS cells.Viral vectors have been used to transfer transcription factors (Oct4, Sox2, c-myc, Klf4, and nanog) to induce reprogramming of mouse fibroblasts, neural stem cells, neural progenitor cells, keratinocytes, B lymphocytes and meningeal membrane cells towards pluripotency. Human fibroblasts, neural cells, blood and keratinocytes have also been reprogrammed towards pluripotency. In this review we have discussed the use of viral vectors for reprogramming both animal and human stem cells. Currently, many studies are also involved in finding alternatives to using viral vectors carrying transcription factors for reprogramming cells. These include using plasmid transfection, piggyback transposon system and piggyback transposon system combined with a non viral vector system. Applications of these techniques have been discussed in detail including its advantages and disadvantages. Finally, current clinical applications of induced pluripotent stem cells and its limitations have also been reviewed. Thus, this review is a summary of current research advances in reprogramming cells into induced pluripotent stem cells.

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Nucleus transfer in mammals: noninvasive approaches for the preparation of cytoplasts

Cited by 42 publications

References 33 publications

Obox4 critically regulates cAMP‐dependent meiotic arrest and MI‐MII transition in oocytes

Obox4 critically regulates cAMP‐dependent meiotic arrest and MI‐MII transition in oocytes

Antimitotic Treatments for Chemically Assisted Oocyte Enucleation in Nuclear Transfer Procedures

Advances in Reprogramming Somatic Cells to Induced Pluripotent Stem Cells

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